Antigenicity and conformational analysis of the Zn2+-binding sites of two Zn2+-metalloproteases: Leishmania gp63 and mammalian endopeptidase-24.11

 
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1996 (EN)
Antigenicity and conformational analysis of the Zn2+-binding sites of two Zn2+-metalloproteases: Leishmania gp63 and mammalian endopeptidase-24.11 (EN)

Soteriadou, K. P. (EN)

Πανεπιστήμιο Ιωαννίνων. Σχολή Θετικών Επιστημών. Τμήμα Χημείας (EL)
Soteriadou, K. P. (EN)

The antigenic properties of the Zn2+-binding region of two Zn2+- metalloproteases, Leishmania surface protease gp63 and mammalian endopeptidase-24.11 (E-24.11), possessing in their active site the characteristic amino acid sequence HEXXH, were investigated by using oligoclonal antibodies raised against two synthetic peptides, V(1)VTHEMAHALGL(11) (pepgp63) and V(1)IGHEITHGFD(11) (pepE-24.11), containing the respective Zn2+-binding sites of the cognate protein. The affinity-purified antibodies, tested on synthetic peptides modelled from the active sites of ten different Zn2+-metalloproteases, showed high selectivity for their respective peptides, However, cross-reactivity was revealed when the antibodies were tested against the gp63 and E-24.11 molecules. A panel of synthetic peptide analogues and peptides of various size was synthesized and used for the fine antigenic characterization of pepgp63 and pepE-24.11. The shortest peptides capable of significant antibody binding were the pentapeptides V(1)VTHE(5) and E(5)ITHG(9) for pepgp63 and pepE-24.11 respectively. His(4) and Glu(5) were found to be indispensable for anti-pepgp63 binding to pepgp63, whereas in the case of pepE-24.11, Glu(5) and His(8) were found to be critical. The conformational characteristics of the two peptides correlate well with the observed differences in their antigenicity. H-1-NMR studies showed that pepgp63 adopts a folded structure whereas pepE-24.11 takes up a rather flexible conformation. Moreover, the antigenically critical His(4) of pepgp63 contributes to the structural stabilization of the peptide. Similarly, the antigenically critical Hiss Of pepE-24.11 is involved in partial structural stabilization of its C-terminal region. The generated antibodies may be useful tools for identifying and classifying proteins possessing similar Zn2+-binding motifs and/or environments. (EN)

nuclear-magnetic-resonance (EN)

Πανεπιστήμιο Ιωαννίνων (EL)
University of Ioannina (EN)

Biochemical Journal (EN)

English

1996

<Go to ISI>://A1996TR39300014

Portland Press (EN)



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